Identifying redox sensors for direct therapy to treat dilated cardiomyopathy

i) Dilated cardiomyopathy (DCM), is a prevalent condition characterised by enlargement and dilation of one or both ventricles, leading to heart failure. Despite its prevalence the prognosis for patients with DCM remains poor, therefore our aim is to identify new strategies for therapy. A key feature of DCM is increased formation of reactive oxygen species (ROS), which can modify susceptible protein thiols through disulphide formation to mediate maladaptive signalling. ii) Therefore, the aim of this study is to identify sites that can be directly targeted for therapy to treat DCM, by mapping proteomics data onto our annotated database of predicted redox-dependent disulphides (ReDisulphID). iii) Here sites of enhanced redox-dependent disulphides in a human iPSC derived DCM model (generated using CRISPR-Cas9 gene editing) will be identified using a new approach for deep coverage and quantitative redox proteomics. Once mapped sites of interest will be characterised using traditional and advanced biochemical, molecular, and cell biology techniques. Here lead targets identified will be assessed in animal and human tissues available within our groups, as well as characterised using CRISPR-Cas9 engineered ‘redox-dead’ human iPSC derived cardiomyocytes. iiv) The aim of year one will be to map potential sites for therapy in a human cell model of DCM using mass spectrometry and bioinformatics. In year two specific sites will be identified for therapy by charactering novel redox-dependent disulphides using CRISPR-Cas9 engineered cells. Finally in year three lead compounds will be identified for therapy to treat DCM, by screening thiol-reactive compound libraries to identify drugs that can be used to selectively regulate disease associated disulphide formation.